The neutral- and basic-amino-acid-transport glycoprotein NBAT (rBAT, D2) expressed in renal and jejunal brush-border membranes interacts with the b0,+AT permease to produce a heteromeric transporter effecting amino acid and cystine absorption. NBAT mutations result in type I cystinuria. The b0,+AT permease is presumed to be the catalytic subunit, but we have been investigating the possibility that cysteine residues within the C-terminus of NBAT are also important for expression of transport function. NBAT mutants were produced with combinations of Cys664/671/683 → Ala substitutions. Mutants with Cys664 → Ala show decreased arginine and cystine transport and specifically lose sensitivity to inhibition of transport by the thiol-group reagent N-ethylmaleimide (NEM). We suggest that the C-terminus of NBAT may have a direct role in the mechanism of System b0,+ transport (the major transport activity defective in type I cystinuria) and that Cys664 of NBAT is the major target for NEM-induced inactivation of the transport mechanism.
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Research Article|
October 24 2000
Cysteine residues in the C-terminus of the neutral- and basic-amino-acid transporter heavy-chain subunit contribute to functional properties of the system b0,+-type amino acid transporter
George J. PETER;
George J. PETER
1Department of Anatomy and Physiology, University of Dundee MSI/WTB Complex, Dundee DD1 5EH, Scotland, U.K.
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Tatiana B. PANOVA;
Tatiana B. PANOVA
1Department of Anatomy and Physiology, University of Dundee MSI/WTB Complex, Dundee DD1 5EH, Scotland, U.K.
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Graham R. CHRISTIE;
Graham R. CHRISTIE
1Department of Anatomy and Physiology, University of Dundee MSI/WTB Complex, Dundee DD1 5EH, Scotland, U.K.
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Peter M. TAYLOR
Peter M. TAYLOR
1
1Department of Anatomy and Physiology, University of Dundee MSI/WTB Complex, Dundee DD1 5EH, Scotland, U.K.
1To whom correspondence should be addressed (e-mail p.m.taylor@dundee.ac.uk).
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Publisher: Portland Press Ltd
Received:
May 17 2000
Revision Received:
July 31 2000
Accepted:
August 01 2000
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2000
2000
Biochem J (2000) 351 (3): 677–682.
Article history
Received:
May 17 2000
Revision Received:
July 31 2000
Accepted:
August 01 2000
Citation
George J. PETER, Tatiana B. PANOVA, Graham R. CHRISTIE, Peter M. TAYLOR; Cysteine residues in the C-terminus of the neutral- and basic-amino-acid transporter heavy-chain subunit contribute to functional properties of the system b0,+-type amino acid transporter. Biochem J 1 November 2000; 351 (3): 677–682. doi: https://doi.org/10.1042/bj3510677
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